• areg
• fgf19
• liver cirrhosis
• liver progenitor cells

• Animals
• Apoptosis/drug effects
• Hepatic Stellate Cells/metabolism
• Rats
• Hepatocytes/metabolism
• Humans
• Liver Cirrhosis/pathology
• Liver Cirrhosis/therapy
• Liver Cirrhosis/metabolism
• Mice
• Male
• Disease Models, Animal
• Stem Cells
• Stem Cell Transplantation
• Rats, Sprague-Dawley

• Cell therapy
• engraftment
• hepatocyte transplantation
• inflammation
• intracellular adhesion molecule 1
• liver injury
• liver regeneration
• niche
• senescence
• senescence associated secretory phenotype

• Blastocyst complementation
• Chronic liver fibrosis
• Hepatic functions
• Hepatocytes
• Interspecies chimeras

• Animals
• Hepatocytes/metabolism
• Hepatocytes/cytology
• Liver Cirrhosis/therapy
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/pathology
• Mice
• Chimera
• Liver/metabolism
• Liver/pathology
• Rats
• Cell Differentiation
• Mice, Inbred C57BL
• Male
• Blastocyst/metabolism
• Blastocyst/cytology
• Chronic Disease
• Cells, Cultured

• high-fat high-fructose diet
• liver fibrosis
• metabolic dysfunction-associated steatotic liver disease
• obesity
• sirtuin

• NA The Morris L. Lichtenstein Jr. Medical Research Foundation

• activin antagonist
• cell cycle arrest
• liver fibrosis
• liver regeneration
• senescence

• Animals
• Humans
• Mice
• Rats
• Activins
• Bromodeoxyuridine
• Fibrosis
• Liver Diseases/drug therapy
• Mice, Inbred C57BL
• Rats, Inbred F344
• Signal Transduction/drug effects

• P30 DK120531 NIDDK NIH HHS
• R01 DK130949 NIDDK NIH HHS
• R01 DK130949 NIH HHS
• P30 DK120531 NIH HHS

• Hnf4a Isoforms
• Liver Epigenomics
• NFκB
• Oncostatin M Receptor (Osmr)
• STAT3

• Animals
• Rats
• Hepatocyte Nuclear Factor 4/genetics
• Hepatocyte Nuclear Factor 4/metabolism
• Hepatocytes/metabolism
• Liver Cirrhosis/pathology
• Liver Failure/metabolism

• R01 CA104292 NCI NIH HHS
• R01 DK090325 NIDDK NIH HHS
• R01 DK099320 NIDDK NIH HHS

• Dlk-1
• cell transplantation
• fetal liver stem/progenitor cells
• human organoids
• iPS cells

• Humans
• Rats
• Animals
• Rats, Inbred F344
• Stem Cell Transplantation/methods
• Induced Pluripotent Stem Cells
• Liver/metabolism
• Hepatocytes/metabolism

• P30 DK120531 NIDDK NIH HHS
• R01 DK130949 NIDDK NIH HHS
• R01 EB028532 NIBIB NIH HHS
• R01 HL141805 NHLBI NIH HHS

Liver fibrosis and hepatocellular carcinoma (HCC) are common adverse consequences of chronic liver injury. The interaction of various risk factors may cause them to happen. Identification of specific biomarkers is of great significance for understanding the occurrence, development mechanisms, and determining the novel tools for diagnosis and treatment of both liver fibrosis and HCC.

To identify liver fibrosis-related core genes, we analyzed the differential expression pattern of core genes in liver fibrosis and HCC.

Gene expression profiles of three datasets, GSE14323, GSE36411, and GSE89377, obtained from the Gene Expression Omnibus (GEO) database, were analyzed, and differentially expressed genes (DEGs) between patients with liver cirrhosis and healthy controls were identified by screening via R software packages and online tool for Venn diagrams. The WebGestalt online tool was used to identify DEGs enriched in biological processes, molecular functions, cellular components, and Kyoto Encyclopedia of Genes and Genomes pathways. The protein–protein interactions of DEGs were visualized using Cytoscape with STRING. Next, the expression pattern of core genes was analyzed using Western blot and immunohistochemistry in a carbon tetrachloride (CCl₄)-induced liver cirrhosis mouse model and in patient liver samples. Finally, Kaplan-Meier curves were constructed using the Kaplan-Meier plotter online server.

Forty-five DEGs (43 upregulated and 2 downregulated genes) associated with liver cirrhosis were identified from three GEO datasets. Ten hub genes were identified, which were upregulated in liver cirrhosis. Western blot and immunohistochemical analyses of the three core genes, decorin (DCN), dermatopontin (DPT), and SRY-box transcription factor 9 (SOX9), revealed that they were highly expressed in the CCl₄-induced liver cirrhosis mouse model. The expression levels of DCN and SOX 9 were positively correlated with the degree of fibrosis, and SOX 9 level in HCC patients was significantly higher than that in fibrosis patients. However, high expression of DPT was observed only in patients with liver fibrosis, and its expression in HCC was low. The gene expression profiling interactive analysis server (GEPIA) showed that SOX9 was significantly upregulated whereas DCN and DPT were significantly downregulated in patients with HCC. In addition, the Kaplan-Meier curves showed that HCC patients with higher SOX9 expression had significantly lower 5-year survival rate, while patients with higher expression of DCN or DPT had significantly higher 5-year survival rates.

The expression levels of DCN, DPT, and SOX9 were positively correlated with the degree of liver fibrosis but showed different correlations with the 5-year survival rates of HCC patients.

• Liver cirrhosis
• Hepatocellular carcinoma
• Bioinformatical analysis
• Decorin
• Dermatopontin
• SRY-box transcription factor 9

• autophagy
• hepatic macrophages
• lipopolysaccharide
• rat
• thioacetamide

Cell therapy provides hope for treatment of advanced liver failure. Proliferating human hepatocytes (ProliHHs) were derived from primary human hepatocytes (PHH) and as potential alternative for cell therapy in liver diseases. Due to the continuous decline of mature hepatic genes and increase of progenitor like genes during ProliHHs expanding, it is challenge to monitor the critical changes of the whole process. Raman microspectroscopy is a noninvasive, label free analytical technique with high sensitivity capacity. In this study, we evaluated the potential and feasibility to identify ProliHHs from PHH with Raman spectroscopy.

Raman spectra were collected at least 600 single spectrum for PHH and ProliHHs at different stages (Passage 1 to Passage 4). Linear discriminant analysis and a two-layer machine learning model were used to analyze the Raman spectroscopy data. Significant differences in Raman bands were validated by the associated conventional kits.

Linear discriminant analysis successfully classified ProliHHs at different stages and PHH. A two-layer machine learning model was established and the overall accuracy was at 84.6%. Significant differences in Raman bands have been found within different ProliHHs cell groups, especially changes at 1003 cm⁻¹, 1206 cm⁻¹ and 1440 cm⁻¹. These changes were linked with reactive oxygen species, hydroxyproline and triglyceride levels in ProliHHs, and the hypothesis were consistent with the corresponding assay results.

In brief, Raman spectroscopy was successfully employed to identify different stages of ProliHHs during dedifferentiation process. The approach can simultaneously trace multiple changes of cellular components from somatic cells to progenitor cells.

The online version contains supplementary material available at 10.1186/s13287-021-02619-9.

• Cell therapy
• Dedifferentiation
• Identification
• Proliferating human hepatocytes
• Raman spectroscopy

• Cell culture techniques
• Cell- and tissue-based therapy
• Rats
• Regenerative medicine
• Tissue engineering
• Transaminases

• Ki-67
• VEGF
• Vinpocetine
• fibrosis
• thioacetamide

• Animals
• Glutathione/metabolism
• Interleukin-6/metabolism
• Liver/drug effects
• Liver/metabolism
• Liver/pathology
• Liver Cirrhosis/chemically induced
• Liver Cirrhosis/drug therapy
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/pathology
• Male
• Malondialdehyde/metabolism
• Nitric Oxide/metabolism
• Oxidative Stress/drug effects
• Protective Agents/pharmacology
• Protective Agents/therapeutic use
• Rats, Sprague-Dawley
• Superoxide Dismutase/metabolism
• Thioacetamide
• Tumor Necrosis Factor-alpha/metabolism
• Vinca Alkaloids/pharmacology
• Vinca Alkaloids/therapeutic use
• Rats

• biliary epithelial cell
• cell therapy
• hepatic progenitor cell
• hepatocyte
• lineage-tracing
• liver regeneration
• plasticity
• reprogramming
• transdifferentiation

• Aptamer
• Artificial organ
• Bioengineering
• Liver tissue engineering
• Re-endothelialization

• AFP, alpha-fetoprotein
• Drug metabolism
• FBS, fetal bovine serum
• Hepatocyte transplantation
• Liver disease modeling
• Liver regeneration
• MACS, magnetic-activated cell sorting
• MEFs, mouse embryonic fibroblasts
• PHHs, primary human hepatocytes
• SHPCs, small hepatocyte progenitor cells
• SULTs, sulfotransferases
• Toxicity testing
• UGTs, glucuronosyl tranferases
• sECs, sinusoidal endothelial cells

• TGF-β1
• development
• differentiation
• fetal hepatic progenitor cells
• fetal liver

• Acute liver failure
• autophagy
• hepatocyte senescence
• liver regeneration
• liver transplantation

According to existing related experiments and research reports, stem cell transplantation therapy has been shown to have a positive effect on the recovery of liver fibrosis/cirrhosis, but for some reason, this therapy still cannot be widely used in clinical work. One of the reasons that cannot be ignored is the low quantity of exogenous stem cells transplanted into the liver in vivo. Thus, we investigated whether the use of the vascular endothelial growth factor (VEGF) can increase the number of stem cell transplants and improve the efficacy of stem cell transplantation therapy.

Using a Sprague-Dawley rat liver fibrosis model, we transplanted into fibrosis liver allograft bone marrow mesenchymal stem cells (BMSCs) which were labelled with chlormethylbenzamido-1,1-dioctadecyl-3,3,3′3′-tetramethylin-docarbocyamine (CM-DiI) or injected VEGF adenovirus solution through the tail vein or conducted the above two operations simultaneously. The cell surface receptor profile of BMSC was examined by flow cytometry and immunofluorescence staining. Hepatic sinusoidal vascular leakage was measured with Evan's blue dye assay. Paraffin section staining, immunofluorescent staining, RT-qPCR (quantitative reverse transcription polymerase chain reaction), and Western blot were used to evaluate hepatic pathological changes and physiology function.

The in vivo study indicated that, comparing with other groups of rats, the rats with combined treatment of BMSC transplantation and VEGF injection exhibited obvious reduction in liver fibrosis. Evan's blue dye assay suggests that after injecting with VEGF adenovirus solution, the rat's hepatic sinusoidal permeability would be increased. We confirmed the expression of very late antigen-4 (VLA4, integrin α₄β₁) on rat BMSCs and the elevated expression of vascular adhesion molecule-1 (VCAM-1) in the hepatic sinusoidal endothelial cells. In addition, the analysis of CM-DiI-labeled BMSCs showed that the BMSC+VEGF group exhibited better cell engraftment and that the engrafted cells were mainly distributed in the hepatic parenchyma. Furthermore, compared with the other situation, it is best to reconstitute the liver secretion and regeneration function of rats after combined application of VEGF and BMSC.

We showed that VEGF promotes the engraftment of BMSCs in liver fibrosis, enhances liver regeneration, and improves liver function. These outcomes may be related to the increasing hepatic sinusoidal endothelium permeability and VCAM-1-increased expression.

The limited proliferative ability of hepatocytes is a major limitation to meet their demand for cell-based therapy, bio-artificial liver device, and drug tests. One strategy is to amplify cells at the hepatoblast (HB) stage. However, expansion of HBs with their bipotency preserved is challenging. Most HB expansion methods hardly maintain the bipotency and also lack functional confirmation.

On the basis of analyzing and manipulating related signaling pathways during HB (derived from human induced pluripotent stem cells, iPSCs) differentiation and proliferation, we established a specific chemically defined cocktails to synergistically regulate the related signaling pathways that optimize the balance of HB proliferation ability and stemness maintenance, to expand the HBs and investigate their capacity for injured liver repopulation in immune-deficient mice.

We found that the proliferative ability progressively declines during HB differentiation process. Small molecule activation of Wnt or inhibition of TGF-β pathways promoted HB proliferation but diminished their bipotency, whereas activation of hedgehog (HH) signaling stimulated proliferation and sustained HB phenotypes. A cocktail synergistically regulating the BMP/WNT/TGF-β/HH pathways created a fine balance for expansion and maintenance of the bipotency of HBs. After purification, colony formation, and expansion for 20 passages, HBs retained their RNA profile integrity, normal karyotype, and ability to differentiate into mature hepatocytes and cholangiocytes. Moreover, upon transplantation into liver injured mice, the expanded HBs could engraft and differentiate into mature human hepatocytes and repopulate liver tissue with restoring hepatocyte mass.

Our data contribute to the understanding of some signaling pathways for human HB proliferation in vitro. Simultaneous BMP/HGF induction, activation of Wnt and HH, and inhibition of TGF-β pathways created a reliable method for long-term stable large-scale expansion of HBs to obtain mature hepatocytes that may have substantial clinical applications.

• Bipotency maintenance
• Expansion
• Hepatoblasts
• Human iPSCs
• Self-renewal

• liver cirrhosis
• liver fibrosis

• Animals
• Disease Models, Animal
• Fetus
• Heterografts
• Humans
• Liver/metabolism
• Liver/pathology
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/pathology
• Liver Cirrhosis/therapy
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Mice, Transgenic
• Stem Cell Transplantation
• Stem Cells/metabolism
• Stem Cells/pathology

• National University Health System (NUHS/NCSP-R)

Matrine (C15H24N2O) is an alkaloid extracted from the Chinese herb Sophora flavescens that has anti-fibrotic and anti-cancer properties. The aim of the present study was to determine the chemopreventive effect of matrine on the development of primary hepatocellular carcinoma (HCC) and its possible association with the suppression of the Notch signaling pathway. The rats were randomly divided into four groups: Control, model, low-dose matrine and high-dose matrine groups. The model was established by combining a partial hepatectomy with diethylnitrosamine (DEN) + 2-acetylaminofluorene (2-AAF). Low- and high-dose matrine groups received intragastric administration of matrine (0.25 and 2.5 g/l of matrine, respectively). DEN + 2-AAF injections and hepatectomy were not performed in the control group. All rats were sacrificed 2, 4 and 7 weeks after hepatectomy. HCC-like histopathological lesions were detected using hematoxylin and eosin staining. The expression levels of α-1-fetoprotein (AFP), albumin (ALB), Notch1 and Hes1 were analyzed using immunohistochemistry. Hepatic lobule structure loss, liver tissue necrosis and inflammatory cell infiltration, and edema degeneration were observed in the model group. By contrast, hepatocyte cord structure was restored and hepatocyte edema degeneration was significantly reduced after 7 weeks of treatment with matrine. In addition, compared with the model group, matrine reduced the expression of AFP, increased the expression of ALB and reduced the expression of Notch1 and Hes1 (only for high-dose matrine; all P<0.05). The findings suggested that matrine could prevent the early development of HCC-like lesions in a rat model, possibly by modulating Notch pathway activation.

• Hes1
• Notch1
• albumin
• hepatocellular carcinoma
• matrine
• α-1-fetoprotein

• R01 DK090325 NIDDK NIH HHS

• copper
• cell therapy
• differentiation
• stem cells
• bone marrow transplantation

• Animals
• Bile Canaliculi
• Bone Marrow/physiology
• Bone Marrow Cells/physiology
• Bone Marrow Transplantation/methods
• Copper/metabolism
• Copper-Transporting ATPases/genetics
• Copper-Transporting ATPases/metabolism
• Copper-Transporting ATPases/physiology
• Dipeptidyl Peptidase 4/metabolism
• Disease Models, Animal
• Female
• Green Fluorescent Proteins/metabolism
• Hepatocytes/metabolism
• Hepatocytes/transplantation
• Hepatolenticular Degeneration/genetics
• Hepatolenticular Degeneration/metabolism
• Hepatolenticular Degeneration/physiopathology
• Liver/metabolism
• Male
• Mice
• Mice, Inbred C57BL
• Myofibroblasts/transplantation

• P30 CA013330 NCI NIH HHS
• P30 DK020541 NIDDK NIH HHS
• P30 DK041296 NIDDK NIH HHS
• R01 DK071111 NIDDK NIH HHS

Many patients with liver disease come to medical attention once they have advanced cirrhosis or acute decompensation. Most often, patients are screened for liver disease via liver function tests (LFTs). There is very limited published data evaluating laboratory values with biopsy-proven stages of hepatic fibrosis. We set out to evaluate whether any correlation exists between routine LFTs and stages of hepatic fibrosis.

A large retrospective observational study on 771 liver biopsies was conducted for evaluating the stage of fibrosis with AST, ALT, INR, BUN, creatinine, platelets, alkaline phosphatase, bilirubin, and albumin. Mean and 95% confidence intervals were used to describe the distributions of serum markers in different fibrosis stages. Multivariable generalized linear models were used and a two-tailed P-value was calculated.

ALT was not statistically significant for any stage, and AST was statistically significant for stage 3 and 4 fibrosis. INR was statistically significant only in stage 4 disease but remained near the upper limit of normal range. Albumin failed to show a clinically relevant association. Platelets remained within normal laboratory range for all stages. The remaining laboratory values failed to show statistical and clinical significance.

The health care burden from chronic liver disease (CLD) will likely continue to rise, unless clinicians are made aware that normal or near normal laboratory findings may be seen in asymptomatic patients. Earlier identification of asymptomatic patients will allow for treatment with new promising modalities and decrease morbidity and mortality from CLD. Our study shows that laboratory values correlate poorly with liver disease.

• HCV
• NAFLD
• chronic liver disease
• cirrhosis
• hepatic fibrosis
• serum markers

• Biomaterials
• Cell sheet
• Cell tracking
• Co-culture
• Differentiation
• Engraftment
• Extracellular matrix
• Hepatic progenitor cells
• Hepatocyte-like cells (HLCs)
• Mesenchymal stem cells (MSCs)

• Animals
• Cell Differentiation/physiology
• Cell- and Tissue-Based Therapy/methods
• Humans
• Liver/cytology
• Liver/physiology
• Liver Diseases/therapy
• Liver Regeneration/physiology
• Liver Transplantation/methods
• Pluripotent Stem Cells/physiology

• MR/K017047/1 Medical Research Council
• MR/K026666/1 Medical Research Council
• MR/L022974/1 Medical Research Council

Although they are expandable in vitro, hepatic progenitors are immature cells and share many immunomarkers with hepatocellular carcinoma, raising potential concerns regarding maltransformation after transplantation. This study investigated the effects of hepatic nuclear factor (HNF) 4α on the proliferation, migration, and maltransformation of hepatic progenitors and determined the feasibility of using these manipulated cells for transplantation.

The effects of HNF4α on rat hepatic progenitors (i.e. hepatic oval cells) were analyzed by HNF4α overexpression and HNF4α shRNA. Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice injured by carbon chloride (CCl₄) were then transplanted with control, HNF4α-overexpressing or HNF4α-suppressing hepatic oval cells. Finally, the engraftment of these cells in the recipient liver was analyzed.

Rat hepatic progenitors (i.e. hepatic oval cells) expressed HNF4α, although less than that in hepatocytes. When HNF4α was overexpressed in these cells, the proliferation and migration of hepatic oval cells were reduced; but when HNF4α was suppressed by shRNA, the proliferation and migration, and even anchorage-independent growth, of these cells were accelerated. RNA microarray and gene functional analysis revealed that suppressing HNF4α not only impaired many biosynthesis and metabolism pathways of hepatocytes but also increased pathways for cancer. When transplanted into CCl₄-injured NOD/SCID mice, few HNF4α-suppressing hepatic oval cells localized into the liver, while control cells and HNF4α-overexpressing cells engrafted into the liver and differentiated into albumin-positive hepatocytes. Interestingly, the hepatocytes derived from HNF4α-overexpressing cells were less migrative and expressed less c-Myc than the cells derived from control cells.

HNF4α constrains proliferation, migration, and maltransformation of hepatic progenitors, and HNF4α-overexpressing hepatic progenitors serve as an optimal candidate for cell transplantation.

The online version of this article (doi:10.1186/s13287-017-0629-8) contains supplementary material, which is available to authorized users.

• Hepatic nuclear factor 4α
• Hepatic progenitors
• Liver repopulation
• Proliferation

• Hepatocyte differentiation
• Heterogeneous cell transplantation
• Liver epithelioid progenitor cells
• Liver fibrosis
• Stem cells therapy

• Cell transplantation
• Epithelial stem/progenitor cells
• FLSPC
• Fetal liver stem/progenitor cells
• Liver repopulation
• Retrorsine
• Thioacetamide induced fibrosis

• cell death
• cell differentiation
• cellular imaging
• fluorescence techniques
• growth factors
• hepatocytes

• Clinical trials
• Fetal hepatocytes
• Hepatocyte transplantation
• Inborn metabolic errors
• Induced pluripotent stem cells
• Liver transplantation
• Neonatal hepatocytes
• Pluripotent stem cells

• hepatocyte
• rat
• repopulation
• transplantation

• CCl4
• cryopreservation
• hepatocyte progenitor cells
• human ES cells
• liver fibrosis

• Bile duct ligation
• Cell transplantation
• Hepatocyte dedifferentiation
• Hepatocyte transdifferentiation
• Osteopontin

• Animals
• Cell Dedifferentiation
• Cells, Cultured
• Dipeptidyl Peptidase 4/analysis
• Hepatocytes/pathology
• Hepatocytes/transplantation
• Liver/pathology
• Liver Cirrhosis, Biliary/pathology
• Phenotype
• Rats
• Rats, Inbred F344

• R01 DK090325 NIDDK NIH HHS

• Activin A
• Smad proteins
• connective tissue growth factor
• liver fibrosis
• oval cell

• Activins/genetics
• Activins/metabolism
• Adult Stem Cells/metabolism
• Animals
• Case-Control Studies
• Connective Tissue Growth Factor/genetics
• Connective Tissue Growth Factor/metabolism
• HEK293 Cells
• Humans
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/pathology
• Rats
• Signal Transduction
• Smad Proteins/metabolism
• Up-Regulation

Cell therapy is an emerging form of treatment for several liver diseases, but is limited by the availability of donor livers. Stem cells hold promise as an alternative to the use of primary hepatocytes. We performed an exhaustive review of the literature, with a focus on the latest studies involving the use of stem cells for the treatment of liver disease. Stem cells can be harvested from a number of sources, or can be generated from somatic cells to create induced pluripotent stem cells (iPSCs). Different cell lines have been used experimentally to support liver function and treat inherited metabolic disorders, acute liver failure, cirrhosis, liver cancer, and small-for-size liver transplantations. Cell-based therapeutics may involve gene therapy, cell transplantation, bioartificial liver devices, or bioengineered organs. Research in this field is still very active. Stem cell therapy may, in the future, be used as a bridge to either liver transplantation or endogenous liver regeneration, but efficient differentiation and production protocols must be developed and safety must be demonstrated before it can be applied to clinical practice.

• bioartificial liver
• cell therapy
• cell transplant
• gene correction
• induced pluripotent stem cells
• liver disease
• regenerative medicine
• stem cell

The Hepatitis B virus genome persists in the nucleus of virus infected hepatocytes where it serves as template for viral mRNA synthesis. Epigenetic modifications, including methylation of the CpG islands contribute to the regulation of viral gene expression. The present study investigates the effects of spontaneous age dependent loss of hepatitis B surface protein- (HBs) expression due to HBV-genome specific methylation as well as its proximate positive effects in HBs transgenic mice.

Liver and serum of HBs transgenic mice aged 5–33 weeks were analyzed by Western blot, immunohistochemistry, serum analysis, PCR, and qRT-PCR.

From the third month of age hepatic loss of HBs was observed in 20% of transgenic mice. The size of HBs-free area and the relative number of animals with these effects increased with age and struck about 55% of animals aged 33 weeks. Loss of HBs-expression was strongly correlated with amelioration of serum parameters ALT and AST. In addition lower HBs-expression went on with decreased ER-stress. The loss of surface protein expression started on transcriptional level and appeared to be regulated epigenetically by DNA methylation. The amount of the HBs-expression correlated negatively with methylation of HBV DNA in the mouse genome.

Our data suggest that methylation of specific CpG sites controls gene expression even in HBs-transgenic mice with truncated HBV genome. More important, the loss of HBs expression and intracellular aggregation ameliorated cell stress and liver integrity. Thus, targeted modulation of HBs expression may offer new therapeutic approaches. Furthermore, HBs-transgenic mice depict a non-infectious mouse model to study one possible mechanism of HBs gene silencing by hypermethylation.

• Animals
• Blotting, Western
• CpG Islands
• DNA Methylation
• Enzyme-Linked Immunosorbent Assay
• Female
• Gene Expression Regulation, Viral
• Gene Silencing
• Hepatitis B
• Hepatitis B Surface Antigens/analysis
• Hepatitis B Surface Antigens/biosynthesis
• Hepatitis B Surface Antigens/blood
• Hepatitis B Surface Antigens/genetics
• Hepatitis B virus/metabolism
• Liver/chemistry
• Liver/virology
• Male
• Mice
• Mice, Transgenic
• Real-Time Polymerase Chain Reaction

Epithelial to mesenchymal transition (EMT), particularly, type 2 EMT, is important in progressive renal and hepatic fibrosis. In this process, incompletely regenerated renal epithelia lose their epithelial characteristics and gain migratory mesenchymal qualities as myofibroblasts. In hepatic fibrosis (importantly, cirrhosis), the process also occurs in injured hepatocytes and hepatic progenitor cells (HPCs), as well as ductular reaction-related bile epithelia. Interestingly, the ductular reaction contributes partly to hepatocarcinogenesis of HPCs, and further, regenerating cholangiocytes after injury may be derived from hepatic stellate cells via mesenchymal to epithelia transition, a reverse phenomenon of type 2 EMT. Possible pathogenesis of type 2 EMT and its differences between renal and hepatic fibrosis are reviewed based on our experimental data.

• animal models
• bile ductular reaction
• epithelial to mesenchymal transition
• hepatic fibrosis
• hepatic progenitor cells
• myofibroblasts
• renal fibrosis

• differentiation
• hepatocyte
• liver disease
• stem cell therapy

AIM: To study the effect of mobilized peripheral blood autologous CD34 positive (CD34⁺) cell infusion in patients with non-viral decompensated cirrhosis.

METHODS: Cirrhotic patients of non-viral etiology were divided into 2 groups based on their willingness to be listed for deceased donor liver transplant (DDLT) (control, n = 23) or to receive autologous CD34⁺ cell infusion through the hepatic artery (study group, n = 22). Patients in the study group were admitted to hospital and received granulocyte colony stimulating factor injections 520 μg/d for 3 consecutive days to mobilize CD34⁺ cells from the bone marrow. On day 4, leukapheresis was done and CD34⁺ cells were isolated using CliniMAC magnetic cell sorter. The isolated CD34⁺ cells were infused into the hepatic artery under radiological guidance. The patients were discharged within 48 h. The control group received standard of care treatment for liver cirrhosis and were worked up for DDLT as per protocol of the institute. Both groups were followed up every week for 4 wk and then every month for 3 mo.

RESULTS: In the control and the study group, the cause of cirrhosis was cryptogenic in 18 (78.2%) and 16 (72.72%) and alcohol related in 5 (21.7%) and 6 (27.27%), respectively. The mean day 3 cell count (cells/μL) was 27.00 ± 20.43 with a viability of 81.84 ± 11.99%. and purity of 80%-90%. Primary end point analysis revealed that at 4 wk, the mean serum albumin in the study group increased significantly (2.83 ± 0.36 vs 2.43 ± 0.42, P = 0.001) when compared with controls. This improvement in albumin was, however, not sustained at 3 mo. However, at the end of 3 mo there was a statistically significant improvement in serum creatinine in the study group (0.96 ± 0.33 vs 1.42 ± 0.70, P = 0.01) which translated into a significant improvement in the Model for End-Stage Liver Disease score (15.75 ± 5.13 vs 19.94 ± 6.68, P = 0.04). On statistical analysis of secondary end points, the transplant free survival at the end of 1 mo and 3 mo did not show any significant difference (P = 0.60) when compared to the control group. There was no improvement in aspartate transaminase, alanine transaminase, and bilirubin at any point in the study population. There was no mortality benefit in the study group. The procedure was safe with no procedural or treatment related complications.

CONCLUSION: Autologous CD 34⁺ cell infusion is safe and effectively improves liver function in the short term and may serve as a bridge to liver transplantation.

• CD34 cell infusion
• Stem cell
• Cirrhosis
• Model for end-stage liver disease
• Liver transplantation

• Adult
• Antigens, CD34/blood
• Biomarkers/blood
• Blood Coagulation
• Female
• Granulocyte Colony-Stimulating Factor/administration & dosage
• Hematopoietic Stem Cell Mobilization
• Hematopoietic Stem Cells/drug effects
• Hematopoietic Stem Cells/immunology
• Hepatic Artery
• Humans
• Immunomagnetic Separation
• India
• International Normalized Ratio
• Leukapheresis
• Liver/blood supply
• Liver/metabolism
• Liver/pathology
• Liver/surgery
• Liver Cirrhosis/blood
• Liver Cirrhosis/diagnosis
• Liver Cirrhosis/mortality
• Liver Cirrhosis/surgery
• Liver Function Tests
• Liver Regeneration
• Liver Transplantation
• Male
• Middle Aged
• Peripheral Blood Stem Cell Transplantation/adverse effects
• Peripheral Blood Stem Cell Transplantation/mortality
• Recovery of Function
• Serum Albumin/metabolism
• Serum Albumin, Human
• Time Factors
• Transplantation, Autologous
• Treatment Outcome

The liver is the largest organ in the body; it has a complex architecture, wide range of functions and unique regenerative capacity. The growing incidence of liver diseases worldwide requires increased numbers of liver transplant and leads to an ongoing shortage of donor livers. To meet the huge demand, various alternative approaches are being investigated including, hepatic cell transplantation, artificial devices and bioprinting of the organ itself. Adult hepatocytes are the preferred cell sources, but they have limited availability, are difficult to isolate, propagate poor and undergo rapid functional deterioration in vitro. There have been efforts to overcome these drawbacks; by improving culture condition for hepatocytes, providing adequate extracellular matrix, co-culturing with extra-parenchymal cells and identifying other cell sources. Differentiation of human stem cells to hepatocytes has become a major interest in the field of stem cell research and has progressed greatly. At the same time, use of decellularized organ matrices and 3 D printing are emerging cutting-edge technologies for tissue engineering, opening up new paths for liver regenerative medicine. This review provides a compact summary of the issues, and the locations of liver support systems and tissue engineering, with an emphasis on reproducible and useful sources of hepatocytes including various candidates formed by differentiation from stem cells.

• Bioartificial liver
• Bioprinting
• Hepatocyte
• Liver
• Liver tissue engineering
• Stem cells